ROMK (Kir 1.1, product of the KCNJ1 gene) channels in the kidney are exquisitely regulated to adjust renal potassium excretion and maintain potassium balance. Clathrin-dependent endocytosis plays a critical role, limiting urinary potassium loss in potassium deficiency. In renal disease, aberrant ROMK endocytosis may contribute to potassium retention and life-threatening hyperkalemia. Available evidence indicates ROMK endocytosis is stimulated by WNKs, kinases that are mutated in Familial Hyperkalemic and Hypertension (FHHt). This application builds on our recent discoveries that: 1) Low potassium directly stimulates ROMK endocytosis in the distal nephron, 2) WNK stimulates ROMK endocytosis by phosphorylating the clathrin-adaptor, ARH, 3) NEDD4 Family of Interacting Proteins, NDFIP1 and 2, interact with ROMK to control post-endocytic processing of the channel. To carry these breakthrough observations toward a completely new understanding of how potassium balance is achieved, we outline plans to: 1) Define the mechanism by which potassium directly regulates ROMK endocytosis; 2) Explore the involvement of this pathway in a mouse model of FHHt, 3) Elucidate the molecular mechanism by which NDFIP1 and NDFIP2 control post-endocytic routing of ROMK to the lysosome. The studies should provide novel insights into the molecular basis of renal K handling and K homeostasis in health and disease while illuminating fundamental mechanisms of membrane protein targeting in the kidney.